A highly specific strategy for in suit detection of DNA with nicking enzyme assisted amplification and lateral flow

Nucleic acid tests are widely applied in clinical diagnosis as well as food safety inspection. The current challenge for nucleic acid tests is to realize in suit detection. Characterized by rapid reaction, amplifications assisted by nicking enzymes are suitable for on-site analysis of nucleic acid. However, a large number of non-specific products are generated, potentially skewing results. Here, we present a visualization method for in suit detection of DNA based on nicking enzyme assisted amplification (NEAA). The method takes advantage of an immunochromatographic strip to ensure the specificity of the results. To avoid non-specific amplification caused by false-priming on the probes, two antigen-modified probes are added at the end of the amplification for immobilizing the specific products on the T-line of the dipsticks. The sensitivity, specificity, and robustness of the method are determined by transgenic soya GTS 40-3-2. No complicated instrument is required during the whole process. The method is also expected to be employed for multiple point-of-care testing in the future. (C) 2017 Elsevier B.V. All rights reserved.